[Analysis of the human papillomavirus in laryngeal squamous cell carcinoma patients].

OBJECTIVE: To analyze the infection of human papillomavirus in laryngeal squamous cell carcinoma patients. METHODS: The pathological samples of 64 clinical diagnosed laryngeal squamous cell carcinoma patients were collected. Lunimex and PCR techniques were used to detect the HPV gene infection and immunohistochemistry method was used to analyze the HPV protein expression in the samples. RESULTS: In the 64 cases, 7 were positive for HPV infection by Luminex and PCR tests. 18 were positive for HPV16/18 E6 protein expression. The total positive rate was about 39. 1%. CONCLUSION: The high HPV infection rate in laryngeal squamous cell carcinoma patients in the study indicated indirectly that the importance of the HPV infection in pathogenesis of the laryngeal squamous cell carcinoma.

Proteomic analysis of differences in barley (Hordeum vulgare) malts with distinct filterability by DIGE.

Filterability is an essential quality parameter of barley malt and significantly impacts productive efficiency and quality of beer. In the study, differences of metabolic capability, rather than of initial contents of macromolecules in barleys, were found to be the main reason for malt filterability gap between the widely used cultivars Dan'er and Metcalfe in China. Comparative proteomics based on fluorescent difference gel electrophoresis (DIGE) was employed to quantitatively analyze proteins of four commercial malts belonging to the two cultivars, and 51 cultivar-differential spots were identified to 40 metabolic proteins by MALDI-TOF/TOF mass spectrometry, mainly including hydrolases and pathogen-related proteins. According to their function analysis and abundance comparison between cultivars, filterability-beneficial and -adverse proteins were putatively proposed. Two most remarkable differential proteins, ß-amylase and serpin Z7, were further investigated to verify their effects on Dan'er malt filterability. These results provide biological markers for barley breeders and maltsters to improve malt filterability. BIOLOGICAL SIGNIFICANCE: To the best of our knowledge, this is the first report of comprehensive investigation of metabolic proteins related to wort filterability of barley malts, and sheds light on clues for filterability improvement. Visible differences in the expression level of metabolic proteins between Dan'er and Metcalfe malts using 2D-DIGE signify a valuable tool for cultivar comparison, illustration of key proteins responsible for filterability and even other qualities of barley malts. And with these explorations on biomarkers of malt filterability and other aspects, there will be higher efficiency and quality of beer brewing, less application of exogenous hydrolases and more expending market for Chinese malting barleys. This article is part of a Special Issue entitled: Translational Plant Proteomics.

Proteome analysis of metabolic proteins (pI 4-7) in barley (Hordeum vulgare) malts and initial application in malt quality discrimination.

Barley malt is essential for beer production. In the present study, the nonprolamin fractions including proteins with structural functions or metabolic activities were extracted from barley malts of the widely used cultivars Gangpi and Baudin in China. The metabolic proteomes (pI 4-7) were constructed and compared using two-dimensional electrophoresis (2DE) followed by matrix-assisted laser desorption/ionization-tandem time-of-flight mass spectrometry (MALDI-TOF/TOF) identification. There were 333 and 354 spots detected in the 2DE gels of Gangpi and Baudin malts, respectively, and about 90% of these spots were shared by the two malts. For all, 377 were successfully identified to 192 proteins, most of which were enzymes and enzyme inhibitors, suggesting important roles in barley malting and the mashing stage of brewing. The Baudin malt was found to contain more spots representing amylases, pathogen-related proteins, and chaperones than the Gangpi malt. In addition, enzymes involved in glycolysis and redox pathways showed significantly different profiles between the two malts, permitting a more in-depth elucidation of the relationship between differential proteins and malt qualities.

[Evaluation of the function of autotransplanted parathyroid tissues].

OBJECTIVE: To evaluate the function autotransplanted parathyroid tissues in thyroid surgery. METHODS: The control group comprised 120 volunteers with normal parathyroid glands in whom serum parathyroid hormone (PTH) levels in blood samples taken from the bilateral elbow head veins. The experimental group comprised 34 patients in whom the parathyroid glands, which were mistakenly cut or could not be preserved during thyroid surgery because of ischemia, were fragmented and autografted into the brachioradialis muscle of the nondominant forearm. On the day of surgery and at 3 days, 7 days, and 2 months postoperatively, contrast imaging and detection of serum PTH levels were performed to evaluate the function of the transplanted parathyroid tissues. RESULTS: In the control group, the mean value of the difference between the bilateral brachiocephalic vein PTH levels was (1.19 ± 0.98) ng/L (maximum 4.52 ng/L). In this study, twice the maximum value (9.04 ng/L) was taken as the effective standard. Based on the 9.04 ng/L effective value, the effective number was 26 patients and the effective rate was 76.5% at 2 months postoperatively in the 34 patients of the experimental group. CONCLUSIONS: The autotransplanted parathyroid tissues showed their functions in most patients, therefore, parathyroid gland autotransplantation is an effective method of preserving function of parathyroid glands that are free or severely ischemic in thyroid surgery.

[Selective neck dissection in clinical node positive low risk papillary thyroid cancer patients].

OBJECTIVE: To explore the improvement of modified radical neck dissection in clinical node positive (cN(+)) low risk papillary thyroid cancer patients. METHODS: Seventy-one cases of papillary thyroid cancer with cN(+) underwent selective neck dissection (IIa, III, IV) from 2007 to 2010 were reviewed, including 10 men and 61 women. All patients were at stage I, including 15 T1, 46 T2, 10 T3. Twenty-six patients were found Positive nodes were found by palpation in 26 cases and by ultrasonagraphy in 45 cases. RESULTS: Cervical lymph node metastasis were confirmed pathologically in 63 cases, with metastatic rate of 88.7%; 100% in cN(+) cases by palpation and 82.2% in cN(+) cases by ultrasonagraphy, respectively. Metastatic nodes existed in one level in 13 cases, two levels in 31 cases, and three levels in 19 cases. Recurrence occurred to 4 patients during follow up, with a recurrent rate of 5.6%, and 2 cases of them were found recurrence in the anterior part of Vb, 2 cases in the carotid sheath. CONCLUSION: Selective neck dissection (IIa, III, IV) is acceptable for cN(+) low risk papillary thyroid cancer patients, which can decrease complications in neck and shoulder greatly.

[The analysis of human papillomavirus infection in lip squamous cell carcinoma patients].

OBJECTIVE: To analyze the infection of human papillomavirus in lip squamous cell carcinoma patients. METHODS: The pathological samples of 9 clinical diagnosed lip cancer patients were collected. Lunimex and PCR techniques were used to detect the HPV gene infection and immunohistochemistry method was used to analyze the HPV protein expression in the samples. RESULTS: In the 9 cases, 1 was positive for HPV16 gene and 7 were positive for HPV16/18 E6 protein expression. The total positive rate was about 8/9. CONCLUSION: The high HPV infection rate in lip cancer patients in the study indicated indirectly that the importance of the direct contact to the infection of HPV what was the basis for pathogenesis of the lip squamous cell carcinoma.

Direct electrochemistry and electrocatalysis of hemoglobin on chitosan-room temperature ionic liquid-TiO(2)-graphene nanocomposite film modified electrode.

TiO(2)-graphene nanocomposite was prepared by hydrolysis of titanium isopropoxide in colloidal suspension of graphene oxide and in situ hydrothermal treatment. The direct electrochemistry and electrocatalysis of hemoglobin in room temperature ionic liquid 1-Butyl-3-methylimidazolium hexafluorophosphate, chitosan and TiO(2)-graphene nanocomposite modified glassy carbon electrode were investigated. The biosensor was examined by using UV-vis spectroscopy, scanning electron microscopy and electrochemical methods. The results indicated that hemoglobin remained its bioactivity on the modified electrode, showing a couple of well-defined and quasi-reversible redox peaks, corresponding to hemoglobin Fe(III)/Fe(II) couple. The kinetic parameters for the electrode reaction, such as the formal potential (E(o')), the electron transfer rate constant (k(s)), the apparent coverage (Γ), and Michaelis-Menten constant (K(m)) were evaluated. The biosensor showed good electrochemical responses to the reduction of H(2)O(2) in the ranges of 1-1170 µM. The detection limit was 0.3 µM (S/N=3). The properties of this composite film, together with the bioelectrochemical catalytic activity, could make them useful in the development of bioelectronic devices, and investigation of electrochemistry of other heme proteins at functional interface.

A graphene-based electrochemical sensor for sensitive determination of caffeine.

An electrochemical sensor based on the electrocatalytic activity of graphene (Gr) for sensitive detection of caffeine is presented. The electrochemical behaviors of caffeine on Nafion-Gr modified glassy carbon electrode (Nafion-Gr/GCE) were investigated by cyclic voltammetry and differential pulse voltammetry. The results showed that the Nafion-Gr/GCE exhibited excellent electrocatalytic activity to caffeine. Caffeine can be effectively accumulated at Nafion-Gr/GCE and produce a sensitive anodic peak. Such electrocatalytic behavior of Gr is attributed to its unique physical and chemical properties, e.g., subtle electronic characteristics and strong adsorptive capability. This electrochemical sensor shows an excellent performance for detecting caffeine with a detection limit of 1.2×10(-7) M (S/N=3), a reproducibility of 5.2% relative standard deviation, and a satisfied recovery from 98.6% to 102.0%. The sensor shows great promise for simple and sensitive determination of caffeine.

Novel electrochemical sensor based on functionalized graphene for simultaneous determination of adenine and guanine in DNA.

A nano-material carboxylic acid functionalized graphene (graphene-COOH) was prepared and used to construct a novel biosensor for the simultaneous detection of adenine and guanine. The direct electrooxidation behaviors of adenine and guanine on the graphene-COOH modified glassy carbon electrode (graphene-COOH/GCE) were carefully investigated by cyclic voltammetry and differential pulse voltammetry. The results indicated that both adenine and guanine showed the increase of the oxidation peak currents with the negative shift of the oxidation peak potentials in contrast to that on the bare glassy carbon electrode. The electrochemical parameters of adenine and guanine on the graphene-COOH/GCE were calculated and a simple and reliable electroanalytical method was developed for the detection of adenine and guanine, respectively. The modified electrode exhibited good behaviors in the simultaneous detection of adenine and guanine with the peak separation as 0.334V. The detection limit for individual determination of guanine and adenine was 5.0×10(-8)M and 2.5×10(-8)M (S/N=3), respectively. Furthermore, the measurements of thermally denatured single-stranded DNA were carried out and the value of (G+C)/(A+T) of single-stranded DNA was calculated as 0.80. The biosensor exhibited some advantages, such as simplicity, rapidity, high sensitivity, good reproducibility and long-term stability.

Label-free amperometric immunobiosensor based on a gold colloid and Prussian blue nanocomposite film modified carbon ionic liquid electrode.

A novel experimental methodology based on a Prussian blue (PB) and gold nanoparticles (AuNPs) modified carbon ionic liquid electrode (CILE) was developed for use in a label-free amperometric immunosensor for the sensitive detection of human immunoglobulin G (HIgG) as a model protein. The CILE was fabricated by using the ionic liquid 1-octyl-3-methylimidazolium hexafluorophosphate as binder. Controllable electrodeposition of PB on the surface of the CILE and coating with 3-aminopropyl triethylene silane (APS) formed a film with high electronic catalytic activity and large surface area for the assembly of AuNPs and further immobilization of HIgG antibody. The electrochemistry of the formed nanocomposite biofilm was investigated by electrochemical techniques including cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. The HIgG concentration was measured through the decrease of amperometric responses in the corresponding specific binding of antigen and antibody. The decreased differential pulse voltammetric values were proportional to the HIgG concentration in two ranges, 0.05-1.25 ng mL(-1) and 1.25-40 ng mL(-1), with a detection limit of 0.001 ng mL(-1) (S/N = 3). This electrochemical immunoassay combined the specificity of the immunological reaction with the sensitivity of the AuNPs, ionic liquid, and PB amplified electrochemical detection and would therefore be valuable for clinical immunoassays.

Direct electrochemistry and electrocatalysis of hemoglobin on carbon ionic liquid electrode.

Hemoglobin in agarose was successfully immobilized on a carbon ionic liquid electrode and the direct electrochemical behavior of hemoglobin was investigated. Room temperature ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate was used as the modifier. Ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy and cyclic voltammetry were used to characterize the hemoglobin on the modified electrode. The results showed that the immobilized hemoglobin retained its bioelectrocatalytic activity. The electrochemistry of hemoglobin provided an opportunity to manufacture a third generation of biosensors. Experimental conditions influencing the biosensor performances such as pH, and potential were optimized and assessed. Under the optimal conditions, hydrogen peroxide was detected in the concentration range from 2x10(-6) to 1.2x10(-3)M with a detection limit of 0.2 microM at S/N=3. The apparent Michaelis-Menten constant was 1.495 mM. The biosensor exhibited some advantages, such as short time respond, high sensitivity, good reproducibility and long-term stability.

Direct electrochemistry of horseradish peroxidase on Nafion/[bmim]PF(6)/agarose composite film modified glassy carbon electrode.

A new strategy to construct electrochemical biosensor for direct electrochemistry of horseradish peroxidase (HRP) on glassy carbon electrode (GCE) based on Nafion, agarose hydrogel and hydrophobic room-temperature ionic liquid (RTIL) 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim]PF(6)) composite as sensing platform has been described. [bmim]PF(6) has good conductivity and wide electrochemical windows and agarose can maintain biological activity well. Nafion/[bmim]PF(6)/agarose composite combines the advantages of [bmim]PF(6) and agarose. Electrochemical impedance spectroscopy (EIS), ultraviolet visible spectroscopy (UV-vis), fourier transform infrared (FT-IR) spectroscopy and cyclic voltammetry (CV) were used to characterize the composite film, showing that the composite film could be effectively constructed on the GCE surface and greatly enhance the electron transfer between HRP and electrode. The factors influencing the performance of the resulting biosensor were studied in detail. The biosensor responded to H(2)O(2) in the linear range from 2x10(-6) to 1.6x10(-4)M with a detection limit of 1.2x10(-7)M (based on the S/N=3). The studied biosensor exhibited good accuracy and high sensitivity. Moreover, the proposed method was economical and efficient.

[Re-operation extent of primary lesion of thyroid cancer].

BACKGROUND AND OBJECTIVE: Many thyroid cancer patients need to receive second operation because of tumor residue. This study was to explore the resection extent of primary lesion in re-operation of thyroid carcinoma. METHODS: Clinical data of 55 thyroid carcinoma patients who received re-operation were reviewed. The patients, including seven men and 48 women, were aged of 18-56. All patients received ipsilateral residual lobectomy, isthmectomy, ipsilateral anterior cervical muscle and level VI lymph node dissection. The optimal extent for radical re-resection of primary lesion was explored. RESULTS: Tumor residue was proved by pathology in 29 patients, with a rate of 52.73%. Of the 29 patients, eight had primary lesion residue, seven had level VI lymph node residue, and 14 had both. Tumor cells were connected with muscle in four patients. Forty-five metastatic lymph nodes were found. CONCLUSION: Ipsilateral residual lobectomy, isthmectomy, ipsilateral anterior cervical muscle and level VI lymph node dissection is the basic re-resection extent of primary lesion for thyroid carcinoma patients.

[Study on the inclusion behavior of cucurbit [7] uril with methylene blue by spectrofluorometric titrations].

The characteristics of host-guest complexation between cucurbit [7] uril (CB7) and methylene blue (MB) were investigated by fluorescence spectrometry in acetate buffer solution at room temperature. It was found that the fluorescence intensity of methylene blue regularly increased upon the addition of cucurbit [7] uril accompanying with a blue-shift of the position of the emission maximum. The results indicate the formation of complex between CB7 and MB at a 1 : 1 complex stoichiometry and the association constant was calculated by applying a deduced equation. 1H NMR spectra were applied to to complement the fluorescence work to verify the formation of the complex. From the temperature dependence of the equilibrium constants, thermodynamics parametersH andS values were obtained, indicating an enthalpic driving force for complexation. The possible interaction mechanism was also discussed. This work may extend the application range of cucurbit [7] uril in biochemistry and pharmaceutical analysis.

[Parotidectomy of deep-lobe tumors].

OBJECTIVE: To evaluate advantages of the superficial lobe preservation in the parotidectomy of deep-lobe tumors, which include much lower incidence in postparotidectomy depression, variable aesthetic deformities, facial nerve injury and Frey's syndrome. METHODS: This retrospective review included 114 patients with benign parotid gland tumors hospitalized from 2001 to 2004, among which 13 patients with deep-lobe parotid tumors and 101 patients in the superficial lobe of parotid gland. Seven in the 13 patients with deep-lobe tumors were operated with the method of the superficial lobe preservation. RESULTS: During median 3 years and 3 months following-up, no any cases with either tumor recurrence, permanent facial nerve injury, postparotidectomy depression, or Frey's syndrome was found in the 7 operated patients. Only 1 case with the facial nerve marginal mandibular branch palsy occurred, but completely recovered over a 3-month period of time. CONCLUSIONS: Preservation of the superficial lobe for deep-lobe tumors parotidectomy could decrease the incidence of parotidectomy complications without any influence in the treatment effect. Additionally, the parotid function preservation and cosmetic appearance after operation also satisfy both the patients and surgeons.

[Analysis of differentially expressed lung metastasis-associated proteins in adenoid cystic carcinoma cell lines].

OBJECTIVE: To analyze differentially expressed metastasis-associated proteins in Adenoid cystic carcinoma cell lines of human salivary gland by proteomics. METHODS: Protein expression alterations of ACC-2 and ACC-M cells were described by 2-D gels. After image analysis by software, proteins of interest were excised from the gels and identified by matrix assisted laser desorption ionization time-of-flight mass spectrometer. RESULTS: 12 protein spots showed significantly differential expression patterns between two cell lines. In the identified protein candidates, transketolase, modulator recognition factor 2, Dim1p homolog, splicing factor (arginine/serine-rich 9) and v-Ha-ras l oncogene were all lowly expressed in the poorly metastatic ACC-2 cell and significantly upregulated in highly metastatic ACC-M cell, while type I collagen pro alpha and tumor necrosis factor (ligand) superfamily member 4 showed a high expression in ACC-2 cells and a low expression in ACC-M cells. Pirin (spot 6) just appears in ACC-2 cell and was not detectable in ACC-M cell, while retinal homeobox protein was just detected in ACC-M cell and did not appear in ACC-2 cell. CONCLUSIONS: The proteins may be involved in the adenoid cystic carcinoma lung metastasis through different mechanisms. Our work may contribute to discover diagnostic markers and therapeutic targets.

Proteomics analysis of differentially expressed metastasis-associated proteins in adenoid cystic carcinoma cell lines of human salivary gland.

Metastasis is the most insidious and life threatening aspect of cancers. However little is known about the molecular mechanisms of tumor metastasis. A poorly metastatic Acc-2 cell line and highly metastatic Acc-M cell line were selected as an experimental model to study on metastatic mechanisms and antimetastatic procedures. In the present study, two-dimensional gel electrophoresis and mass spectrometry are combined to approach the protein profiles associated with tumor metastasis between Acc-2 and Acc-M cell lines. Image analysis of silver stained 2-dimensional gels revealed that 12 protein spots showed significantly quantitative and qualitative variations and mass spectrometry is utilized to further identify these spots. Of the identified proteins, transketolase, Dim1p, v-Ha-ras oncogene, type I collagen pro alpha, tumor necrosis factor (ligand) superfamily member 4, and pirin etc, have shown associations with distinct aspect of tumor metastasis to some extent. The dissimilar expression patterns of these 12 spots indicate the different roles they may play involved in tumor metastasis.